Chloramphenicol is metabolized by a glutathione dependent enzyme system in 100,000 x g supernatant of the rat liver. Two major metabolic fractions are formed, which have been separated by high pressure liquid chromatography. It appears that these products are formed by the hydrolytic dechlorination of chloramphenicol. This pathway of metabolism may represent a general mechanism in which halocarbon compounds are dehalogenates into more water soluble and their excretable products.